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Background Human papillomavirus (HPV) E6/E7 mRNA in situ hybridization (HPV E6/E7 RNAscope) appears to be a sensitive and specific technique in detecting transcriptionally active HPV. We aimed to examine the diagnostic utility of this technique in endocervical adenocarcinoma (ECA), to explore the prognostic factors for ECA patients and develop a clinically useful nomogram based on clinicopathological parameters to predict it. Methods We retrospectively analyzed 200 patients with ECA who had undergone surgery at Sun Yat-sen University Cancer Center from 2010 and 2014. The diagnostic performance of HPV E6/E7 RNAscope were evaluated by receiver operating characteristic (ROC) curve. A prognostic nomogram model including HPV E6/E7 RNAscope was generated based on multivariate Cox regression analysis, then we compared the predictive accuracy of the prognostic model with FIGO staging and treatment using concordance index (C-index), time-dependent ROC (tdROC), and decision curve analysis (DCA). Results The sensitivity and specificity of HPV E6/E7 RNAscope for distinguishing HPV-associated adenocarcinoma (HPVA) from non-HPV-associated adenocarcinoma (NHPVA) in the whole cohort were 75.8% and 80%, respectively. According the univariate analysis and multivariate logistic regression analysis, age, lymphovascular invasion (LVI), lymph node involvement (LNI), and HPV E6/E7 RNAscope were valuable predictive factors for OS. These parameters were incorporated into the nomogram model (nomogram A) compared with FIGO stage and treatment. The C-index of nomogram A for predicting OS was 0.825, which was significantly higher than FIGO stage (C-index = 0.653, p = 0.002) and treatment (C-index = 0.578, p Conclusions HPV E6/E7 RNAscope is highly specific for ECA, and the 4-variable nomogram showed more accurate prognostic outcomes in patients with ECA.

Intraoperative differentiation of primary central nervous system lymphoma (PCNSL) and glioma is of great importance to decision-making for neurosurgeons. However, distinguishing these two diseases based on frozen sections presents a challenge for pathologists. Here, we aim to develop and validate a deep learning model (LGNet) that could accurately differentiate PCNSL from glioma on haematoxylin and eosin (H&E)-stained frozen whole-slide images. In this study, the LGNet was developed and validated to distinguish PCNSL from glioma on independent cohorts, and its performance was compared to that of three pathologists with varying levels of expertise. Additionally, a human-machine fusion approach was designed to consider the diagnostic results from both pathologist and LGNet, to improve the integrative diagnostic performance. A proof of concept study was further evaluated with an online pathological decision support platform. The LGNet achieved high area under the receiver operating characteristic curves (AUROCs) of 0·965 and 0·972 for discriminating PCNSL and glioma on the two external validation cohorts. Moreover, the LGNet outperformed the three pathologists, and assisted them in making the distinction. The diagnostic performance human-machine fusion was further improved using the human-machine fusion. Notably, the performance of LGNet was verified with the proof of concept cohort, and it was shown that the time-consumption of LGNet was significantly less than that of pathologists (P

The metastatic or recurrent potential of localized human papillomavirus-associated endocervical adenocarcinoma (HPVA EAC) is difficult to predict, especially based upon biopsy alone. Recent analyses of small cohorts indicate that high tumor nuclear grade (TNG) and the presence of necrotic tumor debris (NTD) from HPVA EACs in cervical biopsy specimens are highly predictive of nodal metastasis (NM). In the present study, we aimed to investigate how reliably tumoral morphologic features from cervical biopsy specimens predict NM or tumor recurrence (TR) and patient outcomes in a large cohort of endocervical adenocarcinoma patients. A cohort comprised of 397 patients with HPVA EAC treated at 18 institutions was identified, and cervical biopsies were paired with their associated complete tumor resections for a total of 794 specimens. A variety of tumoral histologic features were examined for each paired specimen, including TNG (assessed on a 3-tiered scale of increasing abnormalities-TNG1, TNG2, TNG3) and NTD (defined by the presence of necrotic and apoptotic tumor cells within tumor glandular lumens admixed with granular and eosinophilic amorphous material and inflammatory cells), which were correlated with outcomes. The distribution of TNG in biopsies was as follows: 86 (21.7%) TNG1, 223 (56.2%) TNG2, and 88 (22.2%) TNG3. NTD was identified in 176 (44%) of the biopsy specimens. The sensitivity, specificity, positive predictive value, and negative predictive value of a TNG1 assignment in the biopsy being predictive of the same assignment in the full resection were 0.82 (95% confidence interval [CI]: 0.7-0.9), 0.895 (0.86-0.93), 0.593 (0.48-0.696), and 0.96 (0.94-0.98), respectively. Respective values for an NTD-negative status were 0.89 (95% CI: 0.83-0.92), 0.715 (0.64-0.77), 0.72 (0.65-0.77), and 0.89 (0.83-0.93), respectively. Compared with the other cases in each category, both TNG1 and an NTD-negative status were each significantly associated with lower rates of NM (odds ratio for TNG1=0.245, 95% CI: 0.070-0.857, P=0.0277; for NTD=0.199, 95% CI: 0.094-0.421, P

N6-methyladenosine (m6A) is a highly abundant RNA modification in eukaryotic cells. Methyltransferase-like 3 (METTL3), a major protein in the m6A methyltransferase complex, plays important roles in many malignancies, but its role in cervical cancer metastasis remains uncertain. Here, we found that METTL3 was significantly upregulated in cervical cancer tissue, and its upregulation was associated with a poor prognosis in cervical cancer patients. METTL3 knockdown significantly reduced cervical cancer cell migration and invasion. Conversely, METTL3 overexpression markedly promoted cervical cancer cell metastasis in vitro and in vivo. Furthermore, METTL3 mediated the m6A modification of cathepsin L (CTSL) mRNA at the 5'-untranslated region, and the m6A reader protein insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2) bound to the m6A sites and enhanced CTSL mRNA stability. Our results indicated that METTL3 enhanced CTSL mRNA stability via an m6A-IGF2BP2-dependent mechanism, thereby promoting cervical cancer cell metastasis. These findings provide insights into a novel m6A modification pattern involved in cervical cancer development.

RNA-binding proteins (RBPs) play crucial roles in the post-transcriptional regulation of mRNA during numerous physiological and pathological processes, including tumor genesis and development. However, the role of RNA-binding motif protein 43 (RBM43) in esophageal squamous cell carcinoma (ESCC) has not been reported so far. The current study was the first to evaluate RBM43 protein expression by immunohistochemistry (IHC) in an independent cohort of 207 patients with ESCC, to explore its potential prognostic value and clinical relevance in ESCC. The results indicated that RBM43 protein levels were significantly elevated in ESCC tissues and increased RBM43 expression was associated with age and N categories. In addition, ESCC patients with high expression of RBM43 had shorter overall survival (OS) and disease‐free survival (DFS) than those with low RBM43 expression. Furthermore, when survival analyses were conducted at different clinical stages, overexpression of RBM43 was significantly correlated with shortened survival in patients with ESCC at early stages (TNM stage I–II and N0 stage). Cox regression analysis further proved that high RBM43 expression was an independent predictor of poor prognosis in ESCC patients. In conclusion, increased expression of RBM43 is correlated with malignant attributes to ESCC and predicts unfavorable prognosis, suggesting an effective prognostic biomarker and potential therapeutic target for ESCC.

Micropapillary features are seen in pure mucinous carcinoma of breast (PMC), which is termed mucinous carcinoma with micropapillary features (MPMC). However, whether MPMC can be identified as a morphologically, clinically or genetically distinct entity from PMC remains controversial. In this study, a retrospective review of 161 cases of breast mucinous carcinoma was conducted to assess the clinicopathologic features, prognostic implications, and genomic alterations of MPMC and PMC. MPMCs were identified in 32% of mucinous carcinomas showing an excellent interobserver agreement (ICC = 0.922). MPMCs occurred at a younger age and exhibited higher nuclear grade, more frequent lymph nodal metastasis, lymphovascular invasion, and HER2 amplification compared with PMCs. Survival analyses revealed that MPMCs show decreased progression-free survival compared with PMCs in both unmatched and matched cohorts. A similar outcome of distant disease-free survival was observed only in the unmatched cohort. However, no statistical difference in recurrence score was observed between MPMC and PMC using a 21-gene assay. Notably, both MPMCs and PMCs displayed low mutation burden, common mutations affecting TTN, GATA3, SF3B1, TP53, recurrent 6q14.1-q27 losses, and 8p11.21-q24.3 gains. GATA3, TP53, and SF3B1 were recurrently mutated in MPMCs, while PIK3CA mutations were exclusively detected in PMCs. Moreover, MPMCs harbored 17q and 20q gains as well as 17p losses, while PMCs displayed gains at 6p. PI3K-Akt, mTOR, ErbB, and focal adhesion pathways were more frequently deregulated in MPMCs than in PMCs, which may responsible for the aggressive tumor behavior of MPMCs. Our findings suggest that MPMC is morphologically, clinically, and genetically distinct from PMC.

The “macrotrabecular-massive” (MTM) pattern of hepatocellular carcinoma (HCC) has been suggested to represent a distinct HCC subtype and is associated with specific molecular features. Since the immune microenvironment is heterogenous in HCC, it is important to evaluate the immune microenvironment of this novel variant. CMTM6, a key regulator of PD-L1, is an important immunocheckpoint inhibitor. This study aimed to evaluate the prognostic effect of CMTM6/PD-L1 coexpression and its relationship with inflammatory cells in HCC. We analyzed 619 HCC patients and tumors were classified into MTM and non-MTM HCC subtypes. The expression levels of CMTM6 and PD-L1 in tumor and inflammatory cells were evaluated by immunohistochemistry. The density of inflammatory cells in the cancer cell nest was calculated. Tumoral PD-L1 expression and inflammatory cell density were higher in the MTM type than in the non-MTM type. CMTM6-high expression was significantly associated with shorter OS and DFS than CMTM6-low expression in the whole HCC patient population and the MTM HCC patient population. Moreover, MTM HCC patients with CMTM6/PD-L1 coexpression experienced a higher risk of HCC progression and death. In addition, CMTM6/PD-L1 coexpression was shown to be related to a high density of inflammatory cells. Notably, a new immune classification, based on CMTM6/PD-L1 coexpression and inflammatory cells, successfully stratified OS and DFS in MTM HCC. CMTM6/PD-L1 coexpression has an adverse effect on the prognosis of HCC patients, especially MTM HCC patients. Our study provides evidence for the combination of immune status assessment with anti-CMTM6 and anti-PD-L1 therapy in MTM HCC patients. Electronic supplementary material The online version of this article (10.1007/s00262-020-02691-9) contains supplementary material, which is available to authorized users.

Background: Antibody-dependent cell-mediated cytotoxicity (ADCC) is one of the most important mechanisms of trastuzumab. Fragment C Gamma receptor (FcγR) IIA and IIIA polymorphisms influence the affinity of immunoglobin G (IgG). Recently, FcγRIIA and FcγRIIIA polymorphisms have identified with the efficacy of trastuzumab. However, whether FcγR polymorphisms are associated with the efficacy of trastuzumab in the neoadjuvant setting was unclear. Patients and methods: We retrospectively enrolled 101 patients with HER2-positive breast cancer receiving chemotherapy plus trastuzumab at least four cycles as neoadjuvant therapy and mastectomy in Sun Yat-sen university cancer center from May 2015 to March 2021. Among them, twenty patients were excluded because lacking of blood samples. Polymorphisms of FcγRIIA(rs1801274) and FcγRIIIA(rs396991) were examined by nested polymerase chain reaction (PCR) and sanger sequencing. Lastly, we performed multiple immunohistochemistry (mIHC) to examine the expressions of CD8, CD68, CD57, PD1 and PDL1. Results: Blood samples (n=81) were successfully detected. No significant differences between FcγRIIA/FcγRIIIA genotypes and clinical characteristics, including age, clinical stages, menstrual status, molecular subtyping, treatment and pathological complete response (pCR) rate. In paclitaxel-based treatment subgroup (n=34), FcγRIIIA polymorphism was significantly correlated with pCR rate (P< 0.05,Table1), moreover, the disease-free survivals (DFS) in FcγRIIIA-158V carriers subgroup (high affinity) were significantly longer than that in FcγRIIIA-158 F/F genotype group (low affinity) (P=0.036), while that was not significant difference in patients with anthracycline-based treatment (n=47, P=0.248), indicating that anthracycline enhanced the efficacy of neoadjuvant therapy in patients with FcγRIIIA F/F genotype(low affinity). So far, there is no recurrence events in paclitaxel-based treatment group with FcγRIIIA-158V carriers genotype. The mIHC showed that the ratios of stroma CD8+PD1+ and CD68+PDL1+ cells were significantly higher in anthracycline-based treatment group (Table 2), respectively, indicating that anthracycline improved the efficacy of neoadjuvant targeted therapy by decreasing the ratios of exhausted immune cells. Conclusions: FcγRIIIA-158V genotype is associated with better outcome in HER2-positive breast cancer patients who received paclitaxel combined with trastuzumab as neoadjuvant therapy. Anthracycline improves the outcome of neoadjuvant trastuzumab-treated breast cancer patients with FcγRIIIA F/F genotype by decreasing the ratios of CD8+PD1+ and CD68+PDL1+ cells. Correlation between FcγRIIIA and pCR Correlation between FcγRIIIA and pCR in total population and paclitaxel-based treatment population. Ratios of immune cells Ratios of immune cells in anthracycline-based and paclitaxel-based treatment group. Citation Format: Kaping Lee, Rongzhen Luo, Qianyi Lu, Shusen Wang, Fei Xu. Anthracycline improves the outcome of neoadjuvant trastuzumab-treated breast cancer patients with FcγRIIIA F/F genotype by decreasing the ratios of CD8+PD1+ and CD68+PDL1+ cells [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P5-02-40.

Background Expression of basal-like markers (BMs) is usually observed in triple-negative breast cancer (TNBC) and HER2-overexpressing breast cancer and is related to poor prognosis. Recently, some have reported that BM is expressed in 7.8%-27.1% of HR+/HER2- breast cancers (basal-like phenotype, BM+ BCs), but their biological behaviours remain controversial. This study aimed to compare the differences in clinicopathologic features, clinical outcomes and benefit from adjuvant regimens between BM+ BCs and nonbasal-like phenotype breast cancer (BM- BCs) and evaluate their tumour immune microenvironment.Methods BM+ BCs were defined as EGFR- and/or CK5/6-positive (n=150). Overall, 180 BM- BCs were selected as the control cohort. Clinicopathological characteristics were retrospectively collected. Univariate and multivariate analyses were used to assess the relationship between disease-free survival (DFS), overall survival (OS) and BM status. Stromal tumour-infiltrating lymphocytes (sTILs) were evaluated on haematoxylin & eosin-stained slides, and CD3, CD8, FOPX3, chemokine (C-X-C motif) ligand 13 (CXCL13), CD68, PD-1 and PD-L1 were stained by IHC.Results Compared to BM- BCs, BM+ BCs were more frequently diagnosed at a younger age and had a higher tumour stage, histological grade and necrosis. No obvious differences were seen in lymphovascular invasion or lymph node metastasis. Lower ER and PR expression and higher Ki-67 expression were observed in BM+ BCs. Overall, 62.8% BM+ BCs had p53 mutations. Shorter DFS and OS were related to BM+ BCs (p=0.021 and 0.018, respectively). In multivariate analysis, CK5/6 and clinical stage were independent prognostic factors for DFS and OS. BM+ BCs had a higher proportion of high sTILs with increased CD3, CD8, FOXP3, CXCL13 and CD68 expression. PD-1 and PD-L1 expression was positively related to BM+ BCs.Conclusion Our study demonstrated that more aggressive morphologic features and worse prognosis were associated with BM+ BCs, and its immune-activated status may support new therapeutic strategies, such as immunotherapy, as a potential treatment.

Trophinin-associated protein (TROAP) levels and P53 status have been used to predict the prognosis of various human tumors. However, the clinicopathological and prognostic significance of TROAP expression and P53 status in pulmonary sarcomatoid carcinoma (PSC) has not yet been elucidated. TROAP gene was selected as candidate by bioinformatics analysis based on TCGA and GEO database. We investigated TROAP and P53 expression in 168 cases of surgically resected PSC in three cancer centers using immunohistochemistry (IHC), and then analyzed the correlation between TROAP expression and various clinicopathological parameters, including P53 status and patient prognosis, using several statistical models. The results showed that lung cancer patients with high TROAP mRNA levels had worse prognosis by TCGA and GEO databases. The significant association between the high expression of TROAP, aberrant P53 expression, and adverse overall survival and disease-free survival (P

Epstein–Barr virus-associated gastric cancer (EBVaGC) shows a robust response to immune checkpoint inhibitors. Therefore, a cost-efficient and accessible tool is needed for discriminating EBV status in patients with gastric cancer. Here we introduce a deep convolutional neural network called EBVNet and its fusion with pathologists for predicting EBVaGC from histopathology. The EBVNet yields an averaged area under the receiver operating curve (AUROC) of 0.969 from the internal cross validation, an AUROC of 0.941 on an external dataset from multiple institutes and an AUROC of 0.895 on The Cancer Genome Atlas dataset. The human-machine fusion significantly improves the diagnostic performance of both the EBVNet and the pathologist. This finding suggests that our EBVNet could provide an innovative approach for the identification of EBVaGC and may help effectively select patients with gastric cancer for immunotherapy.

Progestin usage can alter the histologic and cytologic features of HSIL, which may potentially lead to the under-diagnosis of this precancerous lesion. The objective of the present study was to assess the relationship between the usage of progesterone-based contraceptives and the cytologic features of high-grade squamous intraepithelial lesion (HSIL) of the cervix. In this case-control study, we assembled 46 cases of cervical HSIL (CIN 3), including samples from 26 patients with known history of progestin usage (study group) and 20 samples from patients with no history of any exogenous hormone usage (control group). Cell image analysis was performed on all samples using a proprietary software. Immunohistochemical studies for Ki67, p16, estrogen receptor and progesterone receptor were performed on all cases, as was RNA in situ hybridization for HPV subtypes 16 and 18. Compared with the control group cases, the average nuclear size (21.5±1.80 μm) and the nuclear: cytoplasmic ratio (0.28±0.015) of HSILs in the study group cases were significantly smaller (P=0.026) and reduced (P=0.005), respectively. In addition, the study group cases showed reduced nuclear atypia and pleomorphism and significantly reduced mitotic figures (1.74±1.86/mm in study group versus 5.94±1.3/mm in control group, P

Background Endocervical adenocarcinoma (ECA) is further classified as human papillomavirus (HPV)-associated (HPVA) or non-HPVA (NHPVA), per the International Endocervical Adenocarcinoma Criteria and Classification (IECC). HPVA is a glandular tumor with stromal invasion and/or exophytic expansile-type invasion, associated with the typical molecular characteristics of high-risk HPV (HR-HPV) infection. Transforming acidic coiled-coil protein-3 (TACC3),an oncogene that is frequently abnormally expressed,represents a vital biomarker for multiple human malignancies. This study aimed to examine the role of TACC3 in the diagnosis and prognosis of ECA. Methods We analyzed 264 patients with ECA who underwent surgical resection, classifying their tumors into HPVA and NHPVA subtypes. The expression levels of TACC3, P16, MLH1, PMS2, MSH2, MSH6 and Ki-67 in tumors were evaluated by tissue microarray using immunohistochemistry (IHC). HPV subtypes were identified in formalin-fixed paraffin-embedded (FFPE) ECA tissues by the polymerase chain reaction (PCR). Results ECA samples showed increased TACC3 expression relative to adjacent non-carcinoma samples. TACC3 expression was higher in HPVA than in NHPA. In the HPVA subtype, high TACC3 expression was significantly correlated with P16-positive, Ki-67-high expression. Furthermore, TACC3 levels were significantly related to tumor histological type (P = 0.006), nerve invasion (P = 0.003), differentiation (P = 0.004), surgical margin (P = 0.012), parametrium invasion (P = 0.040), P16 expression (P P = 0.004). Additionally, Kaplan–Meier analysis showed that TACC3 upregulation was associated with poor overall survival (OS, P = 0.001), disease-free survival (DFS, P P Conclusions Taken together, our findings identify that TACC3 is a promising complementary biomarker for diagnosis and prognosis for patients with ECA.

Small cell neuroendocrine carcinoma of the uterine cervix (SCNEC) is a rare cancer involving the human papilloma virus (HPV), and has few available treatments. The present work aimed to assess the feasibility of SOX2 and HPV statuses as predictive indicators of SCNEC prognosis.The associations of SOX2 and/or high-risk (HR)-HPV RNA in situ hybridization (RISH) levels with clinicopathological characteristics and prognostic outcomes for 88 neuroendocrine carcinoma (NEC) cases were analyzed.Among these patients with SCNEC, SOX2, P16Overall, the results of the present study suggest that the co-expression of SOX2 with HR-HPV RISH in SCNEC may represent a specific subgroup exhibiting remarkably poorer prognostic outcomes compared with the expression of any one marker alone.

Background Small cell neuroendocrine carcinoma of the uterine cervix (SCNEC) is a rare cancer involving the human papilloma virus (HPV), and has few available treatments. The present work aimed to assess the feasibility of SOX2 and HPV statuses as predictive indicators of SCNEC prognosis. Methods The associations of SOX2 and/or high-risk (HR)-HPV RNA in situ hybridization (RISH) levels with clinicopathological characteristics and prognostic outcomes for 88 neuroendocrine carcinoma (NEC) cases were analyzed. Results Among these patients with SCNEC, SOX2, P16INK4A and HR-HPV RISH expression and SOX2/HR-HPV RISH co-expression were detected in 68(77.3%), 76(86.4%), 73(83.0%), and 48(54.5%), respectively. SOX2-positive and HR-HPV RISH-positive SCNEC cases were associated with poorer overall survival (OS, P = 0.0170, P = 0.0451) and disease-free survival (DFS, P = 0.0334, P = 0.0309) compared with those expressing low SOX2 and negative HR-HPV RISH. Alternatively, univariate analysis revealed that SOX2 and HR-HPV RISH expression, either separately or in combination, predicted the poor prognosis of SCNEC patients. Multivariate analysis revealed that the co-expression of SOX2 with HR-HPV RISH may be an independent factor of OS [hazard ratio = 3.597; 95% confidence interval (CI): 1.085–11.928; P = 0.036] and DFS [hazard ratio = 2.880; 95% CI: 1.199–6.919; P = 0.018] prediction in SCNEC. Conclusions Overall, the results of the present study suggest that the co-expression of SOX2 with HR-HPV RISH in SCNEC may represent a specific subgroup exhibiting remarkably poorer prognostic outcomes compared with the expression of any one marker alone.

Purpose: We aimed to identify the relationship between the genomic characteristics and clinicopathological features of oligo-metastatic breast cancer.Methods: Oligo-metastatic breast cancer diagnosed by pathology from January 2001 and August 2019 were identified and we matched the poly-metastatic patients based on the clinicopathological features of the oligo-metastatic patients included. The database of all genomic alterations was shown according to the FoundationOne CDx reports. Clinicopathological characteristics were collected and the results of next-generation sequencing were analyzed.Results: A total of 26 breast cancer patients were enrolled in our study, including 14 patients with oligo-metastatic disease and 12 patients with poly-metastatic disease. There was no significant difference in number of gene alteration, tumor mutational burden, variants of unknown significance (VUS), and actional mutation in oligo- and poly-metastasis. PIK3CA, TP53 and ERBB2 were the most common shared alterations identified in patients included. Based on the median time of oligo-progression disease (oligo-PD), we divided the patients with oligo-metastasis into longer oligo-PD group (oligo-PD > 31.04 months) and shorter oligo-PD group (oligo-PD ≤ 31.04 months). The analysis of PIK3CA mutation sites showed that H1047R was associated with a good prognosis in patients with metastatic breast cancer. HER2 positive patients with oligo-metastasis was more likely to have a good prognosis. In addition, VUS might also be a potential prognostic biomarker in metastatic breast cancer. Conclusion: Through the genetic analysis of oligo-metastasis, we found PIK3CA H1047R, HER2 and VUS might predict the different clinical outcomes of breast cancer patients with oligo-metastasis for the individualized treatment.

Jin-Ling Duan,1,* Run-Cong Nie,1,2,* Zhi-Cheng Xiang,1,3,* Jie-Wei Chen,3 Min-Hua Deng,1,2 Hu Liang,1,4 Feng-Wei Wang,1 Rong-Zhen Luo,3 Dan Xie,1,3 Mu-Yan Cai1,3 1State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, 510060, People’s Republic of China; 2Department of Surgery, Sun Yat-Sen University Cancer Center, Guangzhou, 510060, People’s Republic of China; 3Department of Pathology, Sun Yat-Sen University Cancer Center, Guangzhou, 510060, People’s Republic of China; 4Department of Nasopharyngeal Carcinoma, Sun Yat-Sen University Cancer Center, Guangzhou, 510060, People’s Republic of China*These authors contributed equally to this workCorrespondence: Mu-Yan Cai; Dan XieState Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-Sen University Cancer Center, Guangzhou, 510060, People’s Republic of ChinaTel +86-20-87343193Fax +86-20-87343170Email caimy@sysucc.org.cn; xiedan@sysucc.org.cnBackground and Aim: To assess the profile of global histone modifications in small hepatocellular carcinoma (small HCC) and identify its prognostic value in predicting recurrence.Methods: The expression profiles of global histone modifications, including H2AK5AC, H2BK20AC, H3K4me2, H3K9AC, H3K18AC, H4K12AC, and H4R3me2, were evaluated with immunohistochemistry in 335 HBV related small HCC patients. Two histone signature classifiers were then developed using least absolute shrinkage and selection operator Cox regression. A nomogram was built using the classifier and independent risk factors. The performances of the classifier and nomogram were assessed by receiver operating characteristic curves.Results: Histone modifications were more pronounced in tumor tissues than in adjacent liver tissues. In tumor tissues, the risk score built based on the seven-histone signature exhibited satisfactory prediction efficiency, with an AUC = 0.71 (0.63– 0.79) for 2-year survival in the training cohort. Patients with a high risk score had shorter recurrence-free survival than those with a low risk score (HR: 1.96, 95% CI: 1.24– 3.08, p = 0.004; HR: 1.95, 95% CI: 1.12– 3.42, p = 0.019; and HR: 1.97, 95% CI: 1.39– 2.80, p < 0.001 for the training, validation and total cohorts, respectively). Furthermore, the statistical nomogram built using the histone classifier for early recurrence had a C-index = 0.68. In non-neoplastic liver tissues, the hepatic signature based on H3K4me2 and H4R3me2 was related to late recurrence (HR: 2.00, 95% CI: 1.15– 3.48, p = 0.01).Conclusion: Global histone modifications in tumor and adjacent liver tissues are novel predictors of early and late recurrence, respectively, in HBV-related small HCC patients.Keywords: small hepatocellular carcinoma, histone modifications, recurrence, LASSO, prognosis

e12607 Background: Antibody-dependent cell-mediated cytotoxicity (ADCC) is one of the most important mechanisms of Trastuzumab. Fragement C Gamma receptor (FcγR) IIA and IIIA polymorphisms influence the affinity of immunoglobin G (IgG). Recently, FcγRIIA and FcγRIIIA polymorphisms have identified with the efficacy of trastuzumab. However, whether FcγR polymorphisms are associated with the efficacy of trastuzumab in the neoadjuvant setting was unclear. Methods: We retrospectively enrolled 101 patients with HER2-positive breast cancer receiving chemotherapy plus trastuzumab as neoadjuvant therapy and mastectomy in Sun Yat-sen university cancer center from May 2015 to March 2021. Among them, twenty patients were excluded because lacking of blood samples. Clinical and pathological characteristics, treatments and outcomes of patients were collected. We purified the genomic DNA from peripheral blood samples, performed nested polymerase chain reaction (PCR) and sanger sequencing to identify the polymorphisms of FcγRIIA(rs1801274) and FcγRIIIA(rs396991). Results: Patient samples (n=81) were successfully detected. No significant differences between FcγRIIA/FcγRIIIA genotypes and clinical characteristics, including age, clinical stage, menstrual status, molecular subtyping, treatment and pathological complete response (pCR) rate. In subgroup analyses, FcγRIIIA polymorphism was significantly correlated with pCR rate in the population who received paclitaxel plus trastuzumab therapy (n=34, P

Background: The metastatic or recurrent potential of clinically localized HPV-associated endocervical adenocarcinoma (HPVA EAC) is difficult to predict. There is a need to develop a practical method that can accurately distinguish apparently localized tumors with a high probability of nodal metastasis (NM) or tumor recurrence (TR) at the initial biopsy phase, where findings can be incorporated into management decisions. Methods: This retrospective study included 794 specimens from 397 patients with primary HPVA EAC treated at 19 institutions. For each patient, data from a paired cervical biopsy (prior to surgery, n=397) and resection specimen, including lymphadenectomy (n=397) were reviewed. A variety of tumoral pathologic/histologic features were examined in each paired-specimen, including tumor nuclear grade (TNG) and necrotic tumor debris (NTD). TNG was assigned on a three-tiered scale of increasing abnormalities (TNG1, TNG2, and TNG3). Any relationships with standard clinicopathologic variables and patient outcomes including NM and TR were evaluated. Findings: Biopsy tumors with low-risk histologic features (TNG1 and NTD-negative) showed markedly better outcomes, with a 0% rate of lymph node metastases or lymphovascular invasion (LVI) and a 1.4% (a single case) rate of tumor recurrence. In contrast, biopsies with TNG 2/3 and/or NTD-positive, showed significantly worse outcomes, with a rate of 19.1% NM, a rate of 12.9% TR, and a rate of 9.6% LVI. The findings remained statistically significant on multivariate analyses. Interpretation: The findings may be of potential clinical utility in the risk stratification of patients with HPVA EACs, and may allow patients with a minimal risk of nodal metastases and tumor recurrence to be identified at the biopsy phase, thereby facilitating more personalized, possibly fertility sparing management options. Funding: Mark and Jane Gibson Endowment fund and department start-up fund, UTSW Medical Center; Natural Science Grant of China and Cervical Cancer Research Award, Zhengzhou, China. Declaration of Interest: None to declare. Ethical Approval: The institutional review board at every participating institution approved this study.

Background To develop and verify pathological models using pathological features basing on hematoxylin and eosin (H&E) images to predict postoperative survival in patients with invasive endocervical adenocarcinoma (ECA). Method: There were 289 patients with ECA classified into training and validation cohorts. A histological signature was produced in 191 patients and verified in the validation group. Histological models combining the histological features were built. They showed increased value compared to the conventional model in terms of individualized prognosis estimation. Results Our model included five selected histological characteristics and was significantly related to overall survival (OS). In the training cohort, it had AUC values of 0.862 and 0.955, respectively, for predicting 3- and 5-year survival; in the validation cohort, the equivalent values were 0.891 and 0.801. In the training cohort, it showed better OS evaluation (C-index: 0.832; 95% confidence interval [CI] = 0.751–0.913) than both the FIGO staging system (C-index: 0.648; 95% CI = 0.542–0.753) and treatment (C-index: 0.687; 95% CI = 0.605–0.769), with advanced efficiency for classifying survival outcomes. In both cohorts, a risk stratification system was built that could precisely stratify patients with stage I and II ECA into high-risk and low-risk subpopulations with significantly different prognoses. Conclusion Our nomogram with five histological signatures had better accuracy in the prediction of OS in patients with ECA. This may contribute to the development of precision medicine in such patients.

Background: Human papillomavirus (HPV) E6/E7 mRNA in situ hybridization (HPV E6/E7 RNAscope) appears to be a sensitive and specific method. We aimed to examine the diagnostic and prognostic utility of this technique in endocervical adenocarcinoma (ECA) and build a useful prognostic nomogram model using this approach. Methods: The model was constructed from a retrospective study of 200 patients with ECA who had undergone surgery at Sun Yat-sen University Cancer Center between 2010 and 2014. The model's predictive efficiency and discriminative capability were defined by a concordance index (C-index) and calibration curve. Results: The overall sensitivity and specificity of HPV E6/E7 RNAscope for distinguishing HPV-associated adenocarcinoma (HPVA) from non HPV-associated adenocarcinoma (NHPVA) in the whole cohort were 75.8% and 80%, respectively, compared to 60.2% and 90.0% for p16 immunohistochemistry, 80.5% and 80.0% for HPV DNA, and 63.3% and 50% for HPV genotype. The independent factors derived from multivariable analysis of the whole cohort to predict overall survival (OS) were age, lymphovascular invasion (LVI), lymph node involvement (LNI), and HPV E6/E7 RNAscope, which were all incorporated into the nomogram with (nomogram B) or without (nomogram A) FIGO stage and treatment. The C-index of nomogram A for predicting OS was 0.825 (p = 0.002 and p < 0.001), which was significantly higher than the C-index for FIGO stage (0.653) and treatment (0.578). No significant difference occurred between nomograms A (0.825) and B (0.836). Furthermore, a risk stratification system was established to accurately stratify patients with ECA into two subgroups with significantly different prognosis. Conclusions: HPV E6/E7 RNAscope is highly specific for ECA, and the proposed nomogram using HPV E6/E7 RNAscope showed more accurate outcome in terms of prognosis in patients with ECA.

BackgroundNeoadjuvant chemotherapy(NACT) is a widely used strategy in the treatment of locally advanced cancers. NACT followed by radical surgery for patients in FIGO stages IB2-IIB has proven to reduce tumor volume and numbers. Under certain conditions, however, NACT followed by radical surgery has not been found to improve overall survival, but rather to accelerate cancer growth and metastasis in current studies.MethodsIn this study, 35 paired pre/post-NACT cervical cancer tissues and 167 cervical cancer samples were examined by immunohistochemical analysis to investigate the role of SLUG in invasion and metastasis. Functional assays and a CHIP assay were performed to determine SLUG’s downstream genes and pathways. A lymph node metastatic mouse model was used to study the effects of SLUG- and MMP3-silencing on the malignant behavior of the cervical cancer cells. ResultsSLUG expression was potentially increased in cervical cancer tissues after neoadjuvant chemotherapy, which in turn positively correlated with pelvic lymph node metastasis. The expression of SLUG following neoadjuvant chemotherapy was identified as a poor survival indicator. SLUG overexpression promoted metastasis in cervical cancer in both in vitro and in vivo models, whereas SLUG silencing had the opposite effect. Mechanically, SLUG is not a prerequisite for EMT activation in cervical cancer models, but contributes to the invasiveness and metastasis of cervical cancer cells by directly tethering to the recognition sequence GCCAGGTGC in the MMP3 promoter region. ConclusionOur results provide mechanistic insight into the potential pro-cancer effect of neoadjuvant chemotherapy, and demonstrates that the SLUG-MMP3 axis could serve as a target for improving the efficacy of chemotherapy.

Centrosomal protein 55 (CEP55) is a centrosome- and midbody-associated protein that is overexpressed in several cancers. However, the underlying molecular mechanism of CEP55-mediated progression and metastasis of esophageal squamous cell carcinoma (ESCC) is not clear. In the current study, we detected CEP55 mRNA by qRT-PCR while protein expression was detected by western blot analysis and immunohistochemistry (IHC). In addition, we knocked down CEP55 and investigated the ability of CEP55 to affect colony formation and migration. Here, we report that CEP55 mRNA and protein expression was significantly increased in ESCC. IHC staining showed that CEP55 expression correlated with TNM stage ( p = 0.046 ) and lymph node metastases ( p = 0.024 ). According to overall survival (OS) and disease-free survival (DFS), patients whose tumors expressed a higher level of CEP55 had a poorer prognosis than those with low expression level of CEP55. A multivariate analysis revealed that CEP55 expression was an independent prognostic indicator for patients with ESCC. Knockdown of CEP55 decreased the colony formation ability and migration of ESCC cells and also reduced the phosphorylation of Src, FAK, and ERK. Therefore, our study implied that CEP55 may be a valuable biomarker and a potential target in the treatment of patients with ESCC.